Compositions and methods for delivery of agents to immune cells

Abstract

The disclosure features immune cell delivery lipid nanoparticle (LNP) compositions that allow for enhanced delivery of agents, e.g., nucleic acids, such as therapeutic and/or prophylactic RNAs, to immune cells, in particular T cells, as well as B cells, dendritic cells and monocytes. The LNPs comprise an effective amount of an immune cell delivery potentiating lipid such that delivery of an agent by an immune cell delivery LNP is enhanced as compared to an LNP lacking the immune cell delivery potentiating agent. Methods of using the immune cell delivery LNPs for delivery of agents, e.g., nucleic acid delivery, for protein expression, for modulating immune cell activity and modulating immune responses are also disclosed.

What is claimed: 
     
         1 . An immune cell delivery lipid nanoparticle comprising:
 (i) an ionizable lipid;   (ii) a sterol or other structural lipid;   (iii) a non-cationic helper lipid or phospholipid;   (iv) an agent for delivery to an immune cell, and   (v) optionally, a PEG-lipid   wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to an immune cell.   
     
     
         2 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises
 a PEG-lipid, and wherein the agent comprises an mRNA encoding a protein of interest.   
     
     
         3 . The immune cell delivery lipid nanoparticle of  claim 1 ,
 wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid or (iii) the non-cationic helper lipid or phospholipid or (v) the PEG lipid is a C1q binding lipid that binds to C1q and/or promotes the binding of the LNP to C1q, as compared to a lipid nanoparticle lacking the C1q binding lipid.   
     
     
         4 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the enhanced delivery is relative to a lipid nanoparticle lacking the immune cell delivery potentiating lipid. 
     
     
         5 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the enhanced delivery is relative to a suitable control. 
     
     
         6 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent stimulates protein expression in the immune cell. 
     
     
         7 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent inhibits protein expression in the immune cell. 
     
     
         8 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent encodes a soluble protein that modulates immune cell activity. 
     
     
         9 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent encodes an intracellular protein that modulates immune cell activity. 
     
     
         10 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent encodes a transmembrane protein that modulates immune cell activity. 
     
     
         11 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent enhances immune function. 
     
     
         12 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the agent inhibits immune function. 
     
     
         13 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the immune cell is a T cell. 
     
     
         14 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the immune cell is a B cell. 
     
     
         15 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the immune cell is a dendritic cell or a myeloid cell. 
     
     
         16 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises a phytosterol, or salt or ester thereof, or a combination of a phytosterol and cholesterol or a salt thereof. 
     
     
         17 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the phytosterol is selected from the group consisting of β-sitosterol, stigmasterol, β-sitostanol, campesterol, brassicasterol, and combinations thereof. 
     
     
         18 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the phytosterol comprises a sitosterol or a salt or an ester thereof. 
     
     
         19 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the phytosterol comprises a stigmasterol or a salt or an ester thereof. 
     
     
         20 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the phytosterol is beta-sitosterol 
       
         
           
           
               
               
           
         
       
       or a salt or an ester thereof. 
     
     
         21 . (canceled) 
     
     
         22 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the immune cell is a T cell and the phytosterol or a salt or ester thereof is selected from the group consisting of β-sitosterol, β-sitostanol, campesterol, brassicasterol, Compound S-140, Compound S-151, Compound S-156, Compound S-157, Compound S-159, Compound S-160, Compound S-164, Compound S-165, Compound S-170, Compound S-173, Compound S-175 and combinations thereof. 
     
     
         23 . The immune cell delivery lipid nanoparticle of  claim 22 , wherein the phytosterol is β-sitosterol. 
     
     
         24 . The immune cell delivery lipid nanoparticle of  claim 22 , wherein the phytosterol is β-sitostanol. 
     
     
         25 . The immune cell delivery lipid nanoparticle of  claim 22 , wherein the phytosterol is campesterol. 
     
     
         26 . The immune cell delivery lipid nanoparticle of  claim 22 , wherein the phytosterol is brassicasterol. 
     
     
         27 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the immune cell is a monocyte or a myeloid cell and the phytosterol or a salt or ester thereof is selected from the group consisting of β-sitosterol, and stigmasterol, and combinations thereof. 
     
     
         28 . The immune cell delivery lipid nanoparticle of  claim 27 , wherein the phytosterol is β-sitosterol. 
     
     
         29 . The immune cell delivery lipid nanoparticle of  claim 27 , wherein the phytosterol is stigmasterol. 
     
     
         30 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises a sterol, or a salt or ester thereof, and cholesterol, wherein the immune cell is a monocyte or a myeloid cell and the sterol or a salt or ester thereof is selected from the group consisting of β-sitosterol-d7, brassicasterol, Compound S-30, Compound S-31 and Compound S-32. 
     
     
         31 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the mol % cholesterol is between about 1% and 50% of the mol % of phytosterol present in the lipid nanoparticle. 
     
     
         32 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the mol % cholesterol is between about 10% and 40% of the mol % of phytosterol present in the lipid nanoparticle. 
     
     
         33 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the mol % cholesterol is between about 20% and 30% of the mol % of phytosterol present in the lipid nanoparticle. 
     
     
         34 . The immune cell delivery lipid nanoparticle of  claim 16 , wherein the mol % cholesterol is about 30% of the mol % of phytosterol present in the lipid nanoparticle. 
     
     
         35 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the ionizable lipid comprises a compound of any of Formulae (I I), (I IA), (I IB), (I II), (I IIa), (I IIb), (I IIc), (I IId), (I IIe), (I IIf), (I IIg), (I III), (I VI), (I VI-a), (I VII), (I VIII), (I VIIa), (I VIIIa), (I VIIIb), (I VIIb-1), (I VIIb-2), (I VIIb-3), (I VIIc), (I VIId), (I VIIIc), (I VIIId), (I IX), (I IXa1), (I IXa2), (I IXa3), (I IXa4), (I IXa5), (I IXa6), (I IXa7), or (I IXa8). 
     
     
         36 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the ionizable lipid comprises a compound selected from the group consisting of Compound X, Compound Y, Compound I-48, Compound I-50, Compound I-109, Compound I-111, Compound I-113, Compound I-181, Compound I-182, Compound I-244, Compound I-292, Compound I-301, Compound I-309, Compound I-317, Compound I-321, Compound I-322, Compound I-326, Compound I-328, Compound I-330, Compound I-331, Compound I-332, Compound I-347, Compound I-348, Compound I-349, Compound I-350, Compound I-352 and Compound I-M. 
     
     
         37 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the ionizable lipid comprises a compound selected from the group consisting of Compound X, Compound Y, Compound I-321, Compound I-292, Compound I-326, Compound I-182, Compound I-301, Compound I-48, Compound I-50, Compound I-328, Compound I-330, Compound I-109, Compound I-111 and Compound I-181. 
     
     
         38 . (canceled) 
     
     
         39 . The immune cell delivery lipid nanoparticle of  claim 37 , wherein immune cell is a T cell and the ionizable lipid comprises a compound selected from the group consisting of Compound I-301, Compound I-321, and Compound I-326. 
     
     
         40 . The immune cell delivery lipid nanoparticle of  claim 36 , wherein immune cell is a monocyte or a myeloid cell and the ionizable lipid comprises a compound selected from the group consisting of Compound X, Compound I-109, Compound I-111, Compound I-181, Compound I-182, and Compound I-244. 
     
     
         41 . The immune cell delivery lipid nanoparticle of  claim 1 , wherein the non-cationic helper lipid or phospholipid comprises a compound selected from the group consisting of DSPC, DPPC, DMPC, DMPE, DOPC, Compound H-409, Compound H-418, Compound H-420, Compound H-421 and Compound H-422. 
     
     
         42 . The immune cell delivery lipid nanoparticle of  claim 41 , wherein the phospholipid is DSPC. 
     
     
         43 . The immune cell delivery lipid nanoparticle of  claim 41 , wherein the immune cell is a T cell and the non-cationic helper lipid or phospholipid comprises a compound selected from the group consisting of DSPC, DMPE, and Compound H-409. 
     
     
         44 .- 46 . (canceled) 
     
     
         47 . The immune cell delivery lipid nanoparticle of  claim 41 , wherein the immune cell is a monocyte or a myeloid cell and the non-cationic helper lipid or phospholipid comprises a compound selected from the group consisting of DOPC, DMPE, and Compound H-409. 
     
     
         48 .- 50 . (canceled) 
     
     
         51 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises a PEG-lipid. 
     
     
         52 . The immune cell delivery lipid nanoparticle of  claim 51 , wherein the PEG-lipid is selected from the group consisting of a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof. 
     
     
         53 . The immune cell delivery lipid nanoparticle of  claim 51 , wherein the PEG lipid comprises a compound selected from the group consisting of Compound P-415, Compound P-416, Compound P-417, Compound P-419, Compound P-420, Compound P-423, Compound P-424, Compound P-428, Compound P-L1, Compound P-L2, Compound P-L3, Compound P-L4, Compound P-L6, Compound P-L8, Compound P-L9, Compound P-L16, Compound P-L17, Compound P-L18, Compound P-L19, Compound P-L22, Compound P-L23 and Compound P-L25. 
     
     
         54 . (canceled) 
     
     
         55 . The immune cell delivery lipid nanoparticle of  claim 51 , wherein the PEG lipid comprises a compound selected from the group consisting of Compound P-428, Compound PL-16, Compound PL-17, Compound PL-18, Compound PL-19, Compound PL-1, and Compound PL-2. 
     
     
         56 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises about 30 mol % to about 60 mol % ionizable lipid, about 0 mol % to about 30 mol % non-cationic helper lipid or phospholipid, about 18.5 mol % to about 48.5 mol % sterol or other structural lipid, and about 0 mol % to about 10 mol % PEG lipid. 
     
     
         57 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises about 35 mol % to about 55 mol % ionizable lipid, about 5 mol % to about 25 mol % non-cationic helper lipid or phospholipid, about 30 mol % to about 40 mol % sterol or other structural lipid, and about 0 mol % to about 10 mol % PEG lipid. 
     
     
         58 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises about 50 mol % ionizable lipid, about 10 mol % non-cationic helper lipid or phospholipid, about 38.5 mol % sterol or other structural lipid, and about 1.5 mol % PEG lipid. 
     
     
         59 . The immune cell delivery lipid nanoparticle of  claim 56 , wherein the mol % sterol or other structural lipid is 18.5% phytosterol and the total mol % structural lipid is 38.5%. 
     
     
         60 . The immune cell delivery lipid nanoparticle of  claim 56 , wherein the mol % sterol or other structural lipid is 28.5% phytosterol and the total mol % structural lipid is 38.5%. 
     
     
         61 . (canceled) 
     
     
         62 . The immune cell delivery lipid nanoparticle of  claim 1 , which comprises:
 (i) about 50 mol % ionizable lipid, wherein the ionizable lipid is a compound selected from the group consisting of Compound I-301, Compound I-321, and Compound I-326;   (ii) about 10 mol % phospholipid, wherein the phospholipid is DSPC;   (iii) about 38.5 mol % structural lipid, wherein the structural lipid is selected from β-sitosterol and cholesterol; and   (iv) about 1.5 mol % PEG lipid, wherein the PEG lipid is Compound P-428.   
     
     
         63 . A method of delivering an agent to an immune cell, the method comprising contacting the immune cell with an immune cell delivery lipid nanoparticle comprising:
 (i) an ionizable lipid;   (ii) a sterol or other structural lipid;   (iii) a non-cationic helper lipid or phospholipid;   (iv an agent for delivery to an immune cell, and   (v) optionally, a PEG-lipid   wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to an immune cell,   such that the agent is delivered to the immune cell.   
     
     
         64 . The method of  claim 63 , wherein the agent comprises a nucleic acid encoding the protein of interest,
 such that expression of the protein of interest is induced in or on the immune cell.   
     
     
         65 . The method of  claim 63 , comprising contacting a T cell with the immune cell delivery lipid nanoparticle,
 wherein the agent comprises a nucleic acid that modulates T cell activation or activity,   such that T cell activation or activity is modulated.   
     
     
         66 . A method of increasing an immune response to a protein, the method comprising contacting immune cells with an immune cell delivery lipid nanoparticle comprising:
 (i) an ionizable lipid;   (ii) a sterol or other structural lipid;   (iii) a non-cationic helper lipid or phospholipid;   (iv) an agent comprising a nucleic acid molecule, and   (v) optionally, a PEG-lipid   wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to immune cells,   such that the immune response to the protein is increased.   
     
     
         67 . A method of increasing a T cell response to a cancer antigen, the method comprising contacting the T cell with an immune cell delivery lipid nanoparticle comprising:
 (i) an ionizable lipid;   (ii) a sterol or other structural lipid;   (iii) a non-cationic helper lipid or phospholipid;   (iv) an agent comprising an mRNA encoding a chimeric antigen receptor (CAR) that binds the cancer antigen, and   (v) optionally, a PEG-lipid   wherein one or more of (i) the ionizable lipid or (ii) the sterol or other structural lipid comprises an immune cell delivery potentiating lipid in an amount effective to enhance delivery of the lipid nanoparticle to immune cells,   such that the T cell response to the cancer antigen is increased.   
     
     
         68 . The method of  claim 66 ,
 wherein the agent comprises an mRNA encoding an antigen of interest,   such that the immune response to the antigen of interest is enhanced in the subject, as compared to the immune response to the antigen of interest induced by an LNP encapsulating the mRNA encoding the antigen of interest but lacking the immune cell delivery potentiating lipid.   
     
     
         69 . The method of  claim 63 , comprising contacting a B cell with the immune cell delivery lipid nanoparticle,
 wherein the agent comprises a nucleic acid molecule that modulates B cell activation or activity,   such that B cell activation or activity is modulated.   
     
     
         70 .- 128 . (canceled)